CN104844511A - Lappaconite hydrobromide compound and preparation thereof - Google Patents

Lappaconite hydrobromide compound and preparation thereof Download PDF

Info

Publication number
CN104844511A
CN104844511A CN201510240193.2A CN201510240193A CN104844511A CN 104844511 A CN104844511 A CN 104844511A CN 201510240193 A CN201510240193 A CN 201510240193A CN 104844511 A CN104844511 A CN 104844511A
Authority
CN
China
Prior art keywords
lappaconitine
dihydrate
compound according
lappaconite
compound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201510240193.2A
Other languages
Chinese (zh)
Other versions
CN104844511B (en
Inventor
朱玉青
徐淑周
徐太平
陈维彩
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shandong Luoxin Pharmaceutical Group Co Ltd
Original Assignee
Shandong Luoxin Pharmaceutical Group Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shandong Luoxin Pharmaceutical Group Co Ltd filed Critical Shandong Luoxin Pharmaceutical Group Co Ltd
Priority to CN201510240193.2A priority Critical patent/CN104844511B/en
Publication of CN104844511A publication Critical patent/CN104844511A/en
Application granted granted Critical
Publication of CN104844511B publication Critical patent/CN104844511B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
    • C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
    • C07D221/22—Bridged ring systems

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention belongs to the field of medicine, and particularly relates to a lappaconite hydrobromide compound and a preparation thereof. The lappaconite hydrobromide compound provided by the invention is lappaconite hydrobromide dihydrate, and the lappaconite hydrobromide dihydrate is measured by using a Cu-K alpha ray to obtain an X-ray powder diffraction pattern (as shown in Figure 1). The invention also discloses a pharmaceutical composition containing the lappaconite hydrobromide dihydrate, and the dosage form of the pharmaceutical composition is an aseptic powder injection, a freeze-dried powder injection or a water injection. The lappaconite hydrobromide dihydrate prepared by the invention is good in stability, especially, the stability under a light condition is improved, and the bioavailability is greatly improved, and the lappaconite hydrobromide dihydrate is more suitable for clinical application.

Description

A kind of Lappaconitine compound and preparation thereof
Technical field
The present invention relates to field of medicaments, specifically, relate to a kind of Lappaconitine compound and preparation thereof.
Background technology
Lappaconitine (C 32h 44n 2o 8.HBr.H 2o) be the hydrobromide monohydrate of the lappaconitine extracted from ranunculaceae plant Aconitum sinomontanum Nakai root, for the non-additive central nervous system analgesia new drug of domestic initiation, it is the moderate anodyne of regulation in Ministry of Health's " cancer patient Three step analgesic ladder governing principle ".Its analgesia intensity is 7 times of pyramidon, suitable with Pethidine analgesic effect, without teratogenesis tire and mutagenesis, and there is toponarcosis, antipyretic and significant resist inflammation on repercussive function of lowering the temperature.The clinical above pain of moderate being mainly used in treatment a variety of causes and causing.Its molecular structural formula is:
At present, about Lappaconitine, some documents and patent are disclosed:
ZL201210001734.2 discloses a kind of pharmaceutical composition of Lappaconitine, and it is the aqueous solution of Lappaconitine, and its pH value is 3.5 ~ 6.0.Be used for the treatment of acute, chronic pharyngitis.
ZL201010574139.9 discloses a kind of process for purification of Lappaconitine, comprise the following steps: the first solvent is joined in Lappaconitine raw material, the weightmeasurement ratio of raw material and the first solvent is made to be 10% ~ 20%, be uniformly mixed, 40 DEG C ~ 70 DEG C heating in water bath 15 minutes ~ 30 minutes, add the second solvent, the second solvent volume is 1/3 ~ 1 times amount of the first solvent volume, leaves standstill to room temperature; Wherein said first solvent is methyl alcohol, and the second solvent is the one in dehydrated alcohol, water, acetone or chloroform; Make its crystallization under putting-20 DEG C ~ 0 DEG C temperature, filter to obtain crystallization; Crystallization is put 30 DEG C ~ 65 DEG C vacuum decompression dryings 1 ~ 4 hour, obtain Lappaconitine.
Patent application 201210102155.7 discloses a kind of Lappaconitine new preparation process, Aconitum sinomontanum Nakai root is pulverized, alkalize with sodium hydroxide, dropping into thermal backflow carries in tank, add the mixed methanol-ethanol solvent 2.7 times (volume) that volume ratio is 2:1, dynamically descend extraction 5 hours 40 DEG C ~ 50 DEG C thermal backflows, obtain crude extract.Again to crude extract carry out acidifying, alkalization separations, crystallization, recrystallizing and refining, Hydrogen bromide react salify obtain finished product.The extract yield of the method is 94%, and content reaches more than 99%.
ZL200410059672.6 discloses a kind of Lappaconitine freeze-dried powder and preparation method thereof.Freeze-dried powder provided by the invention contains Lappaconitine, can also contain pharmaceutically acceptable carrier, its freeze-dried powder good stability, redissolves fast.
Studies have found that, Lappaconitine raw material is to wet or thermally-stabilised, more responsive to light, and Lappaconitine injection liquid of the prior art content after strong illumination is more obvious downtrending, its degradation rate constant is about 2 times of thermostatic accelerated experiment, and pH value of solution change is all not obvious; Lyophilized powder medicament contg and aqueous solution thereof when strong illumination decline obviously.Lappaconitine injection liquid and freeze-dried powder are all unstable to strong illumination.
In order to improve the stability of Lappaconitine in prior art further, and improve the suitability of its clinical application, special proposition the present invention.
Summary of the invention
Primary goal of the invention of the present invention is to propose a kind of Lappaconitine compound.
Second goal of the invention of the present invention is to propose the pharmaceutical composition containing Lappaconitine.
In order to realize object of the present invention, the technical scheme of employing is:
The present invention proposes a kind of Lappaconitine compound, and this Lappaconitine compound is Lappaconitine dihydrate, and its structural formula is such as formula shown in (I):
The X-ray powder diffraction pattern that this Lappaconitine dihydrate uses the measurement of Cu-K alpha-ray to obtain as shown in Figure 1.
The invention still further relates to the preparation method of this Lappaconitine compound, be specially:
(1) Lappaconitine crude product is ground, cross 200 ~ 300 mesh sieves, then join in the anhydrous methanol of 40 ~ 45 DEG C;
(2) water is added while stirring: dimethyl sulfoxide (DMSO): ethyl acetate volume ratio is 1 ~ 4:1 ~ 2:1 ~ 3 mixing solutions, is cooled to 10 ~ 20 DEG C simultaneously;
(3) after mixing solutions adds, leave standstill, continue to be cooled to 0 ~ 4 DEG C, growing the grain 2 ~ 6 hours, washing, dry, obtain Lappaconitine dihydrate.
Wherein: in step (1), prepare the saturated solution of Lappaconitine;
In step (2), stirring velocity is 360 ~ 720 revs/min;
In step (2), the weight of mixed solvent is 8 ~ 12 times of methanol solution weight, and adding speed is 20 ~ 40 ml/min;
In step (3), cooling rate is 1.5 ~ 3 DEG C/h.
The invention still further relates to the pharmaceutical composition containing this Lappaconitine compound, this pharmaceutical composition contains at least one in Lappaconitine dihydrate and pH adjusting agent, stablizer, propping agent, oxidation inhibitor.The formulation of pharmaceutical composition of the present invention is aseptic powder injection, freeze-dried powder or liquid drugs injection.
Below technical scheme of the present invention is made further explanation.
The present invention proposes a kind of Lappaconitine dihydrate, proterties is white crystalline powder, and this hydrate, under Air drying condition, the loss of crystal water can not occur.Lappaconitine of the present invention confirms containing 2 crystal water, and studies as follows it:
1. ultimate analysis
Get the Lappaconitine dihydrate that the present invention prepares and carry out ultimate analysis, adopt U.S. Perkin-Elmer company PE 2,400 II elemental analyser, ultimate analysis (%) measured value: C (54.780), H (6.988), N (4.001), O (22.805), Br (11.385); Conform to the theoretical value of ultimate analysis, ultimate analysis (%) theoretical value is: C (54.778), H (6.985), N (4.000), O (22.804), Br (11.388).
2. differential thermal analysis and thermogravimetric analysis
Get the Lappaconitine dihydrate that the present invention prepares and carry out differential thermal analysis, structure shows, Lappaconitine hydrate of the present invention has endotherm(ic)peak between 140 DEG C ~ 150 DEG C, proves in sample containing crystal water.Its thermogravimetric analysis figure shows the crystal water that its about 140 DEG C lose 2 molecules fast, and without obvious changes in weight before 125 DEG C, confirms that its water molecules lost is crystalline water molecules, but not dissociating water molecule.
3. weight loss on drying and water analysis
Get the Lappaconitine dihydrate that the present invention prepares and be dried to constant weight at 200 DEG C, weightlessness is 5.12%; The weightlessness adopting cassette moisture content tester to measure Lappaconitine dihydrate of the present invention is 5.12%, conforms to theoretical value 5.131%.
4.HPLC purity detecting
Through HPLC purity detecting, the purity of the Lappaconitine dihydrate that the present invention prepares can reach 99.96 ~ 99.98%.
5. crystal formation detects
The X-ray powder diffraction pattern that the Lappaconitine dihydrate that the present invention prepares uses the measurement of Cu-K alpha-ray to obtain as shown in Figure 1.Its X-ray powder diffraction pattern represented with 2 θ ± 0.2 diffraction angle shows characteristic peak at 7.4 °, 11.6 °, 16.0 °, 17.1 °, 20.5 °, 22.0 °, 23.1 °, 25.0 °, 27.1 ° places.
6. fusing point detects
The fusing point of the Lappaconitine dihydrate that the present invention prepares is 227 ~ 228 DEG C.
7. acute toxicity test
Get the Lappaconitine dihydrate that the present invention prepares and carry out acute toxicity test, with C 32h 44n 2o 8.HBr calculate, rat oral LD 50for 48mg/kg, mouse peritoneal injection LD 50for 15.2mg/kg, intravenous injection LD 50for 11.4mg/kg.
Solvent trace (<0.001%) in Lappaconitine dihydrate of the present invention, clinical application is safe and reliable.Confirm through stability test, having good stability of the Lappaconitine hydrate that the present invention prepares, especially improve its stability under illumination condition, be more suitable for clinical application.Lappaconitine dihydrate of the present invention has good mobility, is more adapted to the preparation of preparation.Confirm through acute toxicity test, the toxicity of Lappaconitine dihydrate of the present invention reduces greatly.Find through pharmacokinetic study, the bioavailability of Lappaconitine dihydrate of the present invention improves greatly.
Accompanying drawing illustrates:
Fig. 1 is the X-ray powder diffraction pattern of Lappaconitine dihydrate prepared by embodiment 1;
Fig. 2 is the thermogravimetric analysis figure of the Lappaconitine dihydrate that embodiment 1 prepares;
Fig. 3 is the plasma concentration curve of experimental example 5.
The specific embodiment of the present invention is only limitted to explain further and the present invention is described, does not limit Composition of contents of the present invention.
Embodiment
Embodiment 1
1. Lappaconitine crude product is ground, cross 200 mesh sieves, then join the saturated solution being prepared into Lappaconitine in the anhydrous methanol of 45 DEG C;
2. add water while stirring: dimethyl sulfoxide (DMSO): ethyl acetate volume ratio is 3:1:1 mixing solutions, is cooled to 10 DEG C simultaneously; Stirring velocity is 360 revs/min; The weight of mixed solvent is 8 times of methanol solution weight, and adding speed is 20 ml/min;
3., after mixing solutions adds, leave standstill, continue to be cooled to 2 DEG C, cooling rate is 1.5 DEG C/h; Growing the grain 4 hours, washing, dry, obtain Lappaconitine dihydrate.
This compound crystal detects through high performance liquid chromatography, and purity is 99.97%, yield 94.7%; As shown in Figure 1, thermogravimetric analysis figure as shown in Figure 2 for the X-ray powder diffraction pattern that the measurement of use Cu-K alpha-ray obtains.
Embodiment 2
1. Lappaconitine crude product is ground, cross 300 mesh sieves, then join the saturated solution being prepared into Lappaconitine in the anhydrous methanol of 40 DEG C;
2. add water while stirring: dimethyl sulfoxide (DMSO): ethyl acetate volume ratio is 2:1:2 mixing solutions, is cooled to 20 DEG C simultaneously; Stirring velocity is 720 revs/min; The weight of mixed solvent is 12 times of methanol solution weight, and adding speed is 40 ml/min;
3., after mixing solutions adds, leave standstill, continue to be cooled to 4 DEG C, cooling rate is 1.5 ~ 3 DEG C/h; Growing the grain 3 hours, washing, dry, obtain Lappaconitine dihydrate.
This compound crystal detects through high performance liquid chromatography, and purity is 99.97%, yield 95.1%; As shown in Figure 1, thermogravimetric analysis figure as shown in Figure 2 for the X-ray powder diffraction pattern that the measurement of use Cu-K alpha-ray obtains.
Embodiment 3
1. Lappaconitine crude product is ground, cross 300 mesh sieves, then join the saturated solution being prepared into Lappaconitine in the anhydrous methanol of 42 DEG C;
2. add water while stirring: dimethyl sulfoxide (DMSO): ethyl acetate volume ratio is 3:1:3 mixing solutions, is cooled to 15 DEG C simultaneously; Stirring velocity is 480 revs/min; The weight of mixed solvent is 10 times of methanol solution weight, and adding speed is 30 ml/min;
3., after mixing solutions adds, leave standstill, continue to be cooled to 2 DEG C, cooling rate is 2 DEG C/h; Growing the grain 4 hours, washing, dry, obtain Lappaconitine dihydrate.
This compound crystal detects through high performance liquid chromatography, and purity is 99.97%, yield 94.3%; As shown in Figure 1, thermogravimetric analysis figure as shown in Figure 2 for the X-ray powder diffraction pattern that the measurement of use Cu-K alpha-ray obtains.
Embodiment 4
1. Lappaconitine crude product is ground, cross 300 mesh sieves, then join the saturated solution being prepared into Lappaconitine in the anhydrous methanol of 40 ~ 45 DEG C;
2. add water while stirring: dimethyl sulfoxide (DMSO): ethyl acetate volume ratio is 4:2:3 mixing solutions, is cooled to 12 DEG C simultaneously; Stirring velocity is 720 revs/min; The weight of mixed solvent is 9 times of methanol solution weight, and adding speed is 24 ml/min;
3., after mixing solutions adds, leave standstill, continue to be cooled to 0 DEG C, cooling rate is 3 DEG C/h; Growing the grain 2 hours, washing, dry, obtain Lappaconitine dihydrate.
This compound crystal detects through high performance liquid chromatography, and purity is 99.97%, yield 94.2%; As shown in Figure 1, thermogravimetric analysis figure as shown in Figure 2 for the X-ray powder diffraction pattern that the measurement of use Cu-K alpha-ray obtains.
Embodiment 5: the preparation of aseptic powder injection
Lappaconitine dihydrate embodiment 1 obtained, be that 4:1 mixes with sodium-chlor with mass ratio, aseptically, direct packaging obtains sterile powder injection, and specification is with C 32h 44n 2o 8.HBr 4mg is calculated.
Embodiment 6: the preparation of freeze-dried powder
1. the Lappaconitine dihydrate for preparing of Example 2 is (with C 32h 44n 2o 8.HBr count) 40g, uses water for injection stirring and dissolving, adds N.F,USP MANNITOL 120g, complement to 10L with water for injection;
2. adjust ph is 5.5 ~ 6.0; Add the medicinal carbon that mass percent is 0.01%, adsorb 20 ~ 30 minutes, filtering decarbonization; Essence filter, adds sterilized water for injection to full dose, essence filter;
3. lyophilize: filtrate lyophilize step 2 obtained, is aseptically distributed into 1000, gland, aluminium seal, and to obtain final product.
Lyophilize is divided into pre-freeze, distillation and drying;
Pre-freeze: shelf temperature is down to-20 DEG C with the speed of 3.0 DEG C/min, stops cooling, is incubated 2 hours, then is cooled to-55 DEG C with the speed of 1.5 DEG C/min;
Distillation: be evacuated to 15Pa, rise to-15 DEG C with the speed of 1.5 DEG C/min, be incubated 2 hours; Rise to 15 DEG C with the speed of 3 DEG C/min again to keep 3 hours;
Dry: to rise to 40 DEG C with the speed of 1.2 DEG C/min, dry 2 hours.
Embodiment 7: the preparation of liquid drugs injection
1. the Lappaconitine dihydrate for preparing of Example 2 is (with C 32h 44n 2o 8.HBr count) 40g, join in water for injection, then add sodium-chlor 4g, add to the full amount of water for injection 80%, be heated to 40 DEG C, stirring and dissolving;
2. adjust ph is 5.5 ~ 6.0; Add the medicinal carbon that mass percent is 0.01%, adsorb 20 ~ 30 minutes, filtering decarbonization; Essence filter, adds sterilized water for injection to full dose, essence filter;
3. sterilizing, sealing, packaging, to obtain final product.
Experimental example 1: mobility is tested
The mobility of this experimental example to the Lappaconitine dihydrate of the embodiment of the present invention 1 detects, adopt fixed funnel method, funnel is placed in the suitable height on graph paper, Lappaconitine dihydrate is freely flowed down from flare opening, until the cone top formed contacts with flare opening, measure hypotenuse and the horizontal angle (slope of repose θ) of Lappaconitine dihydrate accumulation horizon.Experimental result is as shown in table 1.
Table 1: mobility experimental result
Batch 1 2 3 4 5 Mean value
θ(°) 25 26 25 24 25 25
From the interpretation of table 1, the mobility of the Lappaconitine dihydrate that the embodiment of the present invention 1 prepares is fine, also detects, obtain similar experimental result to the Lappaconitine dihydrate of other embodiments of the invention.
Experimental example 2: influence factor is tested
1. high temperature test
Three batches 101,102,103 of the Lappaconitine dihydrate that Example 1 prepares, preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, place 10 days at 60 ± 2 DEG C of temperature, in the 5th day and sampling in the 10th day, detect by stability high spot reviews project, test-results compared with 0 day.
2. high humidity test
Three batches 101,102,103 of the Lappaconitine dihydrate that Example 1 prepares, preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, place 10 days under the condition of 25 ± 2 DEG C of relative humidity 90% ± 5%, in the 5th day and sampling in the 10th day, detect by stability high spot reviews project, test-results compared with 0 day.
3. strong illumination test
Three batches 101,102,103 of the Lappaconitine dihydrate that Example 1 prepares, preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, being placed in illumination is place 10 days under the condition of 4500lx, in the 5th day and sampling in the 10th day, detect by stability high spot reviews project, result compared with 0 day.
Influence factor test-results is as shown in table 2.
Table 2:
Result shows: the freeze-dried powder that the Lappaconitine dihydrate that the present invention prepares prepares, and its stability is good, and under high temperature, high humidity, high light conditions, equal retention is stablized.Influence factor experiment is carried out to the Lappaconitine dihydrate that other embodiments of the invention prepare, obtains identical experimental result.
The aseptic powder injection prepared Lappaconitine dihydrate of the present invention, liquid drugs injection carry out influence factor experiment, obtain the experimental result similar to the present embodiment.
Experimental example 3: Acceleration study
Three batches 201,202,203 of the Lappaconitine dihydrate of Example 1 gained, preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, in 42 DEG C, place 6 months under 80%RH condition, at duration of test respectively at 1,2,3,6 sampling at the end of month once, each stability high spot reviews project is tested.Test-results is as shown in table 3.
Table 3:
Result shows: the freeze-dried powder that the Lappaconitine dihydrate that the present invention prepares adopts formula of the present invention and method to prepare, and known through accelerated test result, its stability is good.Acceleration study is carried out to Lappaconitine dihydrate prepared by other embodiment of the present invention, obtains identical experimental result.
The aseptic powder injection prepared Ambroxol HCl hydrate of the present invention, liquid drugs injection carry out influence factor experiment, obtain the experimental result identical with the present embodiment.
Experimental example 4: test of long duration
Three batches 301,302,303 of the Lappaconitine dihydrate of Example 1 gained, preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, place 18 months under temperature 20 DEG C ± 2 DEG C conditions, at duration of test respectively at the 3rd, 6,9,12,18 sampling at the end of month once, each Interventions Requested are tested.Test-results is as shown in table 4:
Table 4:
Result shows: the preparation that the Lappaconitine dihydrate that the present invention prepares adopts formula of the present invention and method to prepare, known through long-term test results, and its stability is good, and equal retention is stablized.Long-term experiment is carried out to Lappaconitine dihydrate prepared by other embodiment of the present invention, obtains identical experimental result.
The aseptic powder injection prepared Lappaconitine dihydrate of the present invention, liquid drugs injection carry out influence factor experiment, obtain the experimental result identical with the present embodiment.
Experimental example 5: simultaneous test
1. strong illumination test
The Lappaconitine monohydrate (comparative example) that the Lappaconitine dihydrate that Example 1 prepares is commercially available, preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, being placed in illumination is place 10 days under the condition of 4500lx, in the 5th day and sampling in the 10th day, detect by stability high spot reviews project, result compared with 0 day.
2. high temperature test
The Lappaconitine dihydrate that Example 1 prepares and commercially available Lappaconitine monohydrate (comparative example), preparation is prepared according to the formula of embodiment 6 and method, simulation listing packaging, put in sealing clean container, place 10 days at 40 ± 2 DEG C of temperature, in the 5th day and sampling in the 10th day, detect by stability high spot reviews project, test-results compared with 0 day.
Comparative test result is as shown in table 2.
Experimental example 6: bioavailability is tested
1. experiment material
1.1 instrument
1100Series LC/MSD Trap SL GC-MS, P230 high pressure constant flow pump, Zorbax SB C 18post (150mm × 4.6mm ID, 5 μm).
2.1.2 medicine and reagent
Lappaconitine reference substance (purity is 99.0%, Gansu Qizheng Tibetan Medicine Group Co., Ltd), AG Virahol, normal hexane, methylene dichloride, hplc grade methanol, heparin sodium, AG acetic acid, nitrogen, sodium hydroxide.
2.1.3 laboratory animal
Male KM mouse, body weight 20 ~ 25g, in whole experimentation, Mouse feeder in the cage of standard, ad lib and drinking-water.
2. experimental technique
1.1 dosage regimen
(l) drug solution preparing
Experimental group: adopt the Lappaconitine dihydrate of embodiment 1 physiological saline configuration 1.0mg/ml solution (with C 32h 44n 2o 8.HBr count);
Control group: adopt commercially available Lappaconitine configuration physiological saline to configure 1.0mg/ml solution;
(2) administration and sample collecting
70 male KM mouse, body weight 20 ~ 25g, duration of test ad lib is drunk water, and is divided into two groups at random.The hydrogen of the experimental group and control group that give 1.0mg/m1 through tail vein injection respectively with the dosage of 2.0mg/kg respectively smells sour lappaconitine solution.Respectively at 1 after administration, 5,10,15,30min and 1,2h get blood 0.5ml through eyeball rear vein beard, puts in heparinised tubes, centrifugal 5min under 12000g, separated plasma.With each time point of dose, process 5 mouse.Sample gives-20 DEG C and is saved to analysis mensuration.
1.2 quantitative analysis method
(l) chromatographic condition
Moving phase is methanol-water-acetic acid (40:60:0.5, v/v/v), and flow velocity is 0.5ml/min, and column temperature is room temperature.
(2) Mass Spectrometry Conditions
Ion source is electron spray ionisation source (ESI source); Source spray voltage is 3.8kV; Dry gas temperature is 325 DEG C; Atomization gas pressure 30psi; Dry gas flow velocity 8l/min; Collision gas (He) pressure of lappaconitine is 0.8Ampl; Collision gas (He) pressure of tetrahydropalmatine is 1.0Ampl; Positive ion mode detects; Scan mode is multiple-reaction monitoring (MRM); Ionic reaction for quantitative analysis is respectively m/z585 → m/z535 (lappaconitine) and m/z356 → m/z192 (tetrahydropalmatine, interior mark).
1.3 plasma sample process
50 μ l inner mark solutions (10.0 μ g/ml tetrahydropalmatine), 100 μ 1 water and 60 μ l sodium hydroxide solutions (1.0mol/l) are added respectively, mixing in 100 μ l mice plasma samples; Add 1ml Extraction solvent (normal hexane: Virahol=95:5, v/v) again, eddy current mixing 1min, reciprocating vibration 10min, centrifugal 5min (3500rpm).After centrifugal, get supernatant liquor and slowly dry up under 40 DEG C of nitrogen gas stream.With 100 μ 1 moving phases redissolution residues, carry out LC/MS/MS analysis.
3. experimental result
Adopt LC/MS/Ms method to mouse vein give hydrogen smell sour lappaconitine after Plasma Concentration measure, the blood concentration-time curve of acquisition is as shown in Figure 3.As seen from Figure 3, under adopting hydrogen of the present invention to smell the plasma concentration curve of sour lappaconitine hydrate, area AUC value is greater than drugs compared, the medicine elimination transformation period t in blood plasma l/2be greater than drugs compared.Illustrate that hydrogen of the present invention smells the bioavailability of sour lappaconitine hydrate higher than prior art.

Claims (9)

1. a Lappaconitine compound, is characterized in that, described Lappaconitine compound is Lappaconitine dihydrate, and its structural formula is such as formula shown in (I):
2. Lappaconitine compound according to claim 1, is characterized in that, the X-ray powder diffraction pattern that described Lappaconitine dihydrate uses the measurement of Cu-K alpha-ray to obtain as shown in Figure 1.
3. Lappaconitine compound according to claim 1, is characterized in that, the preparation method of described Lappaconitine compound is:
(1) Lappaconitine crude product is ground, cross 200 ~ 300 mesh sieves, then join in the anhydrous methanol of 40 ~ 45 DEG C;
(2) water is added while stirring: dimethyl sulfoxide (DMSO): ethyl acetate volume ratio is 1 ~ 4:1 ~ 2:1 ~ 3 mixing solutions, is cooled to 10 ~ 20 DEG C simultaneously;
(3) after mixing solutions adds, leave standstill, continue to be cooled to 0 ~ 4 DEG C, growing the grain 2 ~ 6 hours, washing, dry, obtain Lappaconitine dihydrate.
4. Lappaconitine compound according to claim 3, is characterized in that, in step (1), prepares the saturated solution of Lappaconitine.
5. Lappaconitine compound according to claim 3, is characterized in that, in step (2), stirring velocity is 360 ~ 720 revs/min.
6. Lappaconitine compound according to claim 3, is characterized in that, in step (2), the weight of mixed solvent is 8 ~ 12 times of methanol solution weight, and adding speed is 20 ~ 40 ml/min.
7. Lappaconitine compound according to claim 2, is characterized in that, in step (3), cooling rate is 1.5 ~ 3 DEG C/h.
8. the pharmaceutical composition containing Lappaconitine compound according to claim 1, it is characterized in that, described pharmaceutical composition contains at least one in Lappaconitine dihydrate and pH adjusting agent, stablizer, propping agent, oxidation inhibitor.
9. the pharmaceutical composition of ambroxol compound according to claim 8, is characterized in that, the formulation of described pharmaceutical composition is aseptic powder injection, freeze-dried powder or liquid drugs injection.
CN201510240193.2A 2015-05-12 2015-05-12 A kind of Lappaconitine compound and preparation thereof Active CN104844511B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510240193.2A CN104844511B (en) 2015-05-12 2015-05-12 A kind of Lappaconitine compound and preparation thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510240193.2A CN104844511B (en) 2015-05-12 2015-05-12 A kind of Lappaconitine compound and preparation thereof

Publications (2)

Publication Number Publication Date
CN104844511A true CN104844511A (en) 2015-08-19
CN104844511B CN104844511B (en) 2016-05-04

Family

ID=53844550

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510240193.2A Active CN104844511B (en) 2015-05-12 2015-05-12 A kind of Lappaconitine compound and preparation thereof

Country Status (1)

Country Link
CN (1) CN104844511B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108409657A (en) * 2017-11-24 2018-08-17 孙益民 High-purity lappaconitine and preparation method thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1088781A (en) * 1993-01-01 1994-07-06 中国科学院华南植物研究所 A kind of analgesic and production technology thereof
CN1951922A (en) * 2006-10-20 2007-04-25 佘建文 Lappaconitine hydrobromide preparation method
CN103360311A (en) * 2012-04-10 2013-10-23 广西梧州神农药业有限公司 New preparation process of hydrobromic acid lappaconitine

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1088781A (en) * 1993-01-01 1994-07-06 中国科学院华南植物研究所 A kind of analgesic and production technology thereof
CN1951922A (en) * 2006-10-20 2007-04-25 佘建文 Lappaconitine hydrobromide preparation method
CN103360311A (en) * 2012-04-10 2013-10-23 广西梧州神农药业有限公司 New preparation process of hydrobromic acid lappaconitine

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SUN，WENXIU等: "《X-ray structure analysis of lappaconitine》", 《NATURAL PRODUCT RESEARCH》, vol. 23, no. 10, 29 October 2009 (2009-10-29), pages 960 - 962 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108409657A (en) * 2017-11-24 2018-08-17 孙益民 High-purity lappaconitine and preparation method thereof
CN108409657B (en) * 2017-11-24 2021-08-20 孙益民 High-purity lappaconitine and preparation method thereof

Also Published As

Publication number Publication date
CN104844511B (en) 2016-05-04

Similar Documents

Publication Publication Date Title
Hao et al. Qualitative and quantitative analysis of catechin and quercetin in flavonoids extracted from Rosa roxburghii Tratt
Kong et al. Different pharmacokinetics of the two structurally similar dammarane sapogenins, protopanaxatriol and protopanaxadiol, in rats
US20160310597A1 (en) Caspofungin Acetate Formulations
CN108186559A (en) Cystine injection and its preparation process
CN103833773B (en) A kind of cefathiamidine compound
CN104803860A (en) Ambroxol hydrochloride compound and medicine composition thereof
CN102643255B (en) Andrographolide compound
CN104844511A (en) Lappaconite hydrobromide compound and preparation thereof
CN112574146B (en) Preparation method of butenolide compound composite nanocrystal
CN102836134A (en) Method for preparing mesylate arbidol freeze-dried powder injection preparation
CN104447775B (en) A kind of new alkaloid compound, pharmaceutical composition and its medical usage
CN102670670B (en) Preparation method of ginkgo dipyridolum injection with high content of ginkgo terpene lactones
Sun et al. In vitro and in vivo evaluation of the influences of polysaccharides derived from Glycyrrhiza uralensis on three alkaloids and potential interaction mechanisms
CN103159710B (en) Antiviral decalin derivate
Guo et al. Pharmacokinetic study on hyperoside in Beagle's dogs
CN101081213B (en) Butane diacid(5-androstene-17-ketone- 3beta -hydroxyl group ) diester solid dispersoid and method for making same and applications thereof
CN111233877A (en) Galanthamine pamoate and preparation method thereof
CN104910144A (en) Tropisetron hydrochloride compound and preparation thereof
CN103479617A (en) Dehydroandrographolide derivative composition and preparation method as well as application thereof
CN109232764A (en) A kind of rhizoma menispermi polysaccharide and its preparation method and application
CN103265459B (en) A kind of novel sodium houttuyfonate compound, its preparation method and pharmaceutical composition thereof
CN113195484B (en) Crystal form of propionamide derivative and preparation method thereof
Chen et al. An LC‐MS/MS method for determination of 3, 6′‐disinapoylsucrose in rat plasma and its application to a pharmacokinetic study
Solomun et al. Methylprednisolone and its related substances in freeze-dried powders for injections
CN105017241B (en) A kind of aztreonam compound and its preparation

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
EXSB Decision made by sipo to initiate substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant